Recovery of cytomegalovirus DNA from newborn saliva samples by different methods

Oran Goshen; David M Goldfarb; Laura Book; Peter Tilley; Soren Gantt

doi:10.1016/j.jcv.2018.05.001

Recovery of cytomegalovirus DNA from newborn saliva samples by different methods

J Clin Virol. 2018 Jul:104:73-76. doi: 10.1016/j.jcv.2018.05.001. Epub 2018 May 5.

Authors

Oran Goshen¹, David M Goldfarb², Laura Book³, Peter Tilley⁴, Soren Gantt⁵

Affiliations

¹ British Columbia Children Hospital Research Institute,University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.; Departments of Pediatrics, University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.. Electronic address: oran.goshen@gmail.com.
² British Columbia Children Hospital Research Institute,University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.; Pathology and Laboratory Medicine, University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.. Electronic address: david.goldfarb@cw.bc.ca.
³ British Columbia Children Hospital Research Institute,University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.; Pathology and Laboratory Medicine, University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.. Electronic address: lbook@cw.bc.ca.
⁴ British Columbia Children Hospital Research Institute,University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.; Pathology and Laboratory Medicine, University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.. Electronic address: ptilley@cw.bc.ca.
⁵ British Columbia Children Hospital Research Institute,University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.; Departments of Pediatrics, University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.; Pathology and Laboratory Medicine, University of British Columbia, 4480 Oak Street, V6H 3N1, Canada.. Electronic address: sgantt@cfri.ca.

PMID: 29753934
DOI: 10.1016/j.jcv.2018.05.001

Abstract

Background: Polymerase chain reaction (PCR) testing of saliva swabs has been shown to be an attractive method to screen for congenital cytomegalovirus (CMV) infection. However, few data are available regarding the impact of pre-analytic methods on CMV DNA recovery from saliva samples under various conditions.

Objectives: The purpose of this study was to evaluate the impact of various transport media and transport conditions on the recovery of CMV DNA from spiked and clinical saliva samples from newborns using PCR testing.

Study design: Negative control saliva samples were spiked with known concentrations of World Health Organization CMV standards. Collection was simulated using filter paper and different types of swabs which were stored either dry or in various transport media at specified temperatures. Specimens were tested by quantitative PCR at multiple time points. Results were compared with samples taken from newborns infected with CMV using the same methods.

Results: Nucleic acid stabilization media demonstrated superior CMV stability for up to 3 weeks. Also observed was a reduction in the quantity of spiked CMV DNA over time in swab samples stored at room temperature without media or in conventional universal transport media. Swab samples from newborns infected with CMV did not demonstrate loss of CMV DNA after a week of room temperature storage in nucleic acid stabilization media or conventional universal transport media.

Conclusions: Although nucleic acid preservation media appear to enhance CMV DNA recovery from spiked control samples over prolonged transport times and diverse conditions, this differential yield appears be to less significant for actual clinical samples collected from infected newborns. Further evaluation of pre-analytic conditions using actual clinical samples is warranted.

Keywords: CMV; Congenital; Cytomegalovirus; PCR; Saliva; Screening.

Publication types

Evaluation Study

MeSH terms

Adult
Cytomegalovirus / genetics
Cytomegalovirus / isolation & purification*
Cytomegalovirus Infections / congenital*
Cytomegalovirus Infections / diagnosis*
DNA, Viral / analysis
DNA, Viral / genetics
DNA, Viral / isolation & purification*
Humans
Infant, Newborn
Real-Time Polymerase Chain Reaction / methods
Saliva / virology*
Specimen Handling / methods*
Temperature
Time Factors
Young Adult

Substances

DNA, Viral